Bailey A M, Mena G L, Herrera-Estrella L
Departamento de Ingeniería Genética de Plantas, Centro de Investigación y de Estudios Avanzados (CINVESTAV), Guanajuato, Mexico.
Curr Genet. 1993 Jan;23(1):42-6. doi: 10.1007/BF00336748.
Phytophthora capsici, P. citricola, P. cinnamomi and P. citrophthora were transformed without the removal of cell walls by particle acceleration with plasmids containing the beta-glucuronidase gene and hygromycin B resistance. Transformants were detected by histochemical and fluorometric beta-glucuronidase assays and confirmed by Southern-blot hybridization. It was found that the promoter of a plant virus is functional in Phytophthora. In addition, a method was designed to visually identify homogeneous transformed colonies, derived from zoospores of transformed multinucleated Phytophthora mycelia, based on blue color development on plates containing X-Gluc.
利用含有β-葡萄糖醛酸酶基因和潮霉素B抗性的质粒，通过粒子加速法对辣椒疫霉、柑橘褐腐疫霉、肉桂疫霉和柠檬疫霉进行了不脱细胞壁的转化。通过组织化学和荧光β-葡萄糖醛酸酶分析检测转化体，并通过Southern杂交进行确认。发现植物病毒的启动子在疫霉中具有功能。此外，还设计了一种方法，基于在含有X-Gluc的平板上产生蓝色，从转化的多核疫霉菌丝体的游动孢子中直观地鉴定出均匀的转化菌落。
